Dental composition comprising chelator and base

ABSTRACT

Non-toxic consumable compositions and formulations having chelator and base are disclosed with synergistic effects on microbial metabolism and/or growth and/or pathogenic effectors and their use to promote and maintain health in mammals. The non-toxic consumable compositions have more than one chelator and/or more than one base. Methods for selecting said chelator and base composition and methods for detecting conditions in which selected compositions may be used are also disclosed. The synergistic compositions and methods can be used for maintaining health, promoting health and treating diseases.

CROSS-REFERENCE TO RELATED APPLICATION

This application is the U.S. national phase of PCT Application No.PCT/US2014/024613 filed on Mar. 12, 2014, which claims the benefit ofU.S. Provisional Patent Application No. 61/965,678 filed on Feb. 5, 2014and U.S. Provisional Patent Application No. 61/851,748 filed on Mar. 12,2013, the disclosures of which are incorporated in their entirety byreference herein.

BACKGROUND OF THE INVENTION

Microbes play a central role in both mammalian health and disease, withcertain microbes playing a central role in host physiology and healthwhile other microbes promote or exacerbate disease. Health-promoting andpathogenic microbes may include bacteria, fungi, parasites and othermicrobes and may be found in biofilms.

Microbial biofilms are aggregates of microbial cells that adhere to eachother and to a surface. The adherent cells are frequently embeddedwithin a self-produced matrix, also referred to as an extracellularmatrix generally composed of extracellular nucleic acids, proteins andpolysaccharides (commonly dextran). Biofilms have been demonstrated toexist on biological or inanimate surfaces in home, agricultural,industrial and healthcare settings. A problem is often encountered withbiofilms harboring pathogenic microbes and their toxic effectors thatpose a health risk.

It is recognized that antibiotics have multiple limitations in theirability to inhibit the pathogenic effects of biofilms in home,agricultural, industrial and healthcare settings. Numerous attempts toneutralize the pathogenic effects of biofilms including behavioralchanges, procedural changes, anti-microbial coatings and next generationantibiotics, have been made with limited success and pose significanttoxicity risk to mammals. Non-toxic and effective compositions andmethods of use are needed to limit the adverse effects of microbes andmicrobial biofilms with limited disruption of the healthy microbialflora.

Throughout this specification, various patents, patent applications andother types of publications (e.g., journal articles) are referenced. Thedisclosure of all patents, patent applications, and publications citedherein are hereby incorporated by reference in their entirety for allpurposes.

BRIEF SUMMARY OF THE INVENTION

The invention provided herein discloses, inter alia, non-toxiccompositions comprising a synergistic amount of chelator and base forlimiting the adverse effects of microbes and microbial biofilms while atthe same time, limiting disruption of healthy microbial flora in anindividual.

Accordingly, in one aspect, provided herein are compositions comprisinga chelator and a base, wherein the chelator and base provide microbialaffecting activity. In some embodiments, the chelator is selected fromthe group consisting of siderophores, natural chelators and syntheticchelators. In another embodiment, the chelator is cyclodextrin orcyclodextrin derivative, ferrichrome, citrate, EDTA, EGTA, pectin ormodified pectin. In another embodiment, more than one chelator is used.In another embodiment, the concentration of chelator is between 0.0005%and 30% of the composition. In another embodiment, the base is selectedfrom the group consisting of natural bases, synthetic bases and agentshaving basic properties. In another embodiment, the base is KOH, NaOH,pyridoxal-5-phosphate, pyridoxamine, pyridoxine, vitamin K, lysine,arginine, lysozyme, alpha-galactase, tris amine or sodium bicarbonate.In another embodiment, more than one base is used. In anotherembodiment, the concentration of base is between 0.0001% and 15% of thecomposition. In another embodiment, the composition further comprises anenhancer. In another embodiment, the enhancer comprises proline,phenylalanine, boric acid, ascorbic acid or extracts from citrus,berries, teas, peppermint, mint or cinnamon. In another embodiment, theconcentration of the enhancer is between 0.0001% and 10% of thecomposition. In another embodiment, the composition is for oral care. Inanother embodiment, the base comprises sodium bicarbonate andpyridoxal-5-phosphate and the chelator comprises alpha-cyclodextrin. Inanother embodiment, the composition is formulated in a tablet, capsule,rapid melt tablet, thin strip, gum or mouthwash. In another embodiment,the composition administered at least once per day. In anotherembodiment, the composition is used to promote oral health, treat orprevent cavities, periodontitis, halitosis and gingivitis. In anotherembodiment, the composition is used following consumption of aphosphoric acid or ascorbic acid-containing beverage. In anotherembodiment, the composition is used following consumption of food.

In another aspect, provided herein are methods of promoting oral healthin an individual comprising: (i) identifying an individual at increasedrisk for developing or exhibiting signs of oral disease, (ii) measuringpH, leukocyte esterase, nitrate, microbial marker levels, orplaque-staining dye retention in the mouth of the individual, (iii)comparing said levels to healthy levels to determine if oral pathologyis present, and (iv) if evidence of oral pathology is present,contacting an effective amount of a composition comprising a chelatorand a base to the oral surface. In some embodiments, said method isrepeated on a daily, a monthly, quarterly or annual basis. In someembodiments, said composition comprises EDTA or alpha-cyclodextrin andsodium bicarbonate or pyridoxal-5-phosphate. In some embodiments, theeffective amount of EDTA and alpha-cyclodextrin is between 0.0001% and30% of the composition. In some embodiments, the effective amount ofsodium bicarbonate and pyridoxal-5-phosphate is between 0.0001% and 15%of the composition. In some embodiments, oral pathology is indicated bythe individual having an oral pH below 6.2. In some embodiments, theindividual is determined to have oral pathology based on the presence ofone or more of the following: halitosis, tooth plaque, tooth decay, or acavity. In some embodiments, the individual is determined to have oralpathology based on the presence of Streptococcus mutans.

In further aspects, provided herein are methods for limiting oreliminating microbes and/or microbial biofilms in an oral cavity of anindividual comprising contacting a surface in the oral cavity with aneffective amount of a composition comprising a chelator and a base. Insome embodiments, said method is repeated on a daily, a monthly,quarterly or annual basis. In some embodiments, said compositioncomprises EDTA or alpha-cyclodextrin and sodium bicarbonate orpyridoxal-5-phosphate. In some embodiments, the effective amount of EDTAand alpha-cyclodextrin is between 0.0001% and 30% of the composition. Insome embodiments, the effective amount of sodium bicarbonate andpyridoxal-5-phosphate is between 0.0001% and 15% of the composition. Insome embodiments, the method further comprises assessing oral pathologyin the individual, wherein oral pathology is indicated by the individualhaving an oral pH below 6.2. In some embodiments, the method furthercomprises assessing the individual for oral pathology based on thepresence of one or more of: halitosis, tooth plaque, tooth decay, and/ora cavity. In some embodiments, the method further comprises assessingthe individual for the presence of Streptococcus mutans.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A and 1B depict the effect of agents with increasing baseproperties +/−chelator on microbial growth.

FIG. 2 depicts the effect of individual agents vs. synergistic effect ofagent combinations on biofilms.

FIG. 3 depicts the effect of individual agents vs. agent combinations onmicrobial acid production.

FIGS. 4A and 4B depict the effect of compositions and formulations onhuman oral health markers.

FIGS. 5A and 5B depict an exemplary differential effect on pathogenicand commensal microbes.

FIG. 6 depicts an exemplary effect of chelator and basicity of agents onmicrobial growth.

FIG. 7 depicts an exemplary effect of chelator and base compositions onfungal growth.

FIGS. 8A, 8B, and 8C depict an exemplary effect of chelator and basecompositions on food biofilms.

DETAILED DESCRIPTION

The present invention generally relates to non-toxic compositionscomprising a synergistic amount of chelator and base, whereby said basemay comprise one or more hydroxyl group, pyridine ring, nitrogen groupor amine group and whereby said chelator may comprise one or more ringedstructure capable of chelating metal, lipid, volatile aromatic compoundor microbial component. Said compositions may comprise a chelator, abase and an enhancer, whereby said enhancer enhances the utility of thechelator or base. Said compositions may be used to limit microbialgrowth and unwanted metabolic activity of microbes by contacting asurface to formulations comprising synergistic chelator and basecompositions. Said compositions may be used to selectively inhibit thegrowth and viability of certain microbes while maintaining the viabilityof many commensal microbes of the flora. Said compositions may be usedto selectively alter the metabolism of certain microbes. Saidcompositions may be used to prevent or treat conditions where microbesinduce host inflammation. Said compositions may comprise consumableformulations or may comprise topical formulations. In certain aspects,said compositions may be administered to biological and inanimatesurfaces comprising a microbial biofilm. Non-toxic compositions of thecurrent invention may be applied frequently to a biological or inanimatesurface to promote oral care, skin care, food safety, overall health andsurface care. Non-toxic compositions may be used in products for oralcare, skin care, food safety, overall health and surface care.

I. Definitions

As used herein, the singular form “a”, “an”, and “the” includes pluralreferences unless indicated otherwise.

It is understood that aspects and embodiments of the invention describedherein include “comprising,” “consisting,” and “consisting essentiallyof” aspects and embodiments.

An “individual” can be a vertebrate, a mammal, or a human. Mammalsinclude, but are not limited to, farm animals, sport animals, pets,primates, mice and rats. Individuals also include companion animalsincluding, but not limited to, dogs and cats. In one aspect, anindividual is a human.

An “effective amount” or “therapeutically effective amount” refers to anamount of therapeutic compound, such as a composition comprising achelator and a base, administered to an individual, either as a singledose or as part of a series of doses, which is effective to produce adesired therapeutic effect.

A “clinician” or “medical researcher” or “veterinarian” as used herein,can include, without limitation, doctors, nurses, physician assistants,lab technicians, research scientists, clerical workers employed by thesame, or any person involved in determining, diagnosing, aiding in thediagnosis or influencing the course of treatment for the individual.

It is intended that every maximum numerical limitation given throughoutthis specification includes every lower numerical limitation, as if suchlower numerical limitations were expressly written herein. Every minimumnumerical limitation given throughout this specification will includeevery higher numerical limitation, as if such higher numericallimitations were expressly written herein. Every numerical range giventhroughout this specification will include every narrower numericalrange that falls within such broader numerical range, as if suchnarrower numerical ranges were all expressly written herein.

II. Compositions of the Invention

Disclosed herein are non-toxic compositions, comprising chelator andbase, for use in limiting microbial growth and unwanted metabolicactivity and pathogenic effectors of microbes on surfaces. Saidcompositions may be applied to a biological or inanimate surface.Surfaces may comprise biological surfaces such as food, dermal surfaces,tooth surfaces, mucosal surfaces, gut surfaces, ocular surfaces or otherand may comprise inanimate surfaces such as objects, floors, counters,utensils, handles, or other.

Also disclosed herein are non-toxic consumable compositions, comprisingchelator and base, for use in limiting microbial growth, biofilmproduction and unwanted metabolic activity and pathogenic effectors ofmicrobes involved in mammalian disease. Many diseases have an underlyingmicrobial or microbial-induced inflammatory component that contributesto disease initiation or progression. Both gram-positive andgram-negative bacteria contain several components within theirmembranes, such as lipopolysaccharide (endotoxin), lipoteichoic acid,CpG DNA, flagella, pili, and other antigenic molecules that are potentagonists of Toll-like receptors (TLRs), which induces pro-inflammatorycytokine secretion of epithelial cells and immune cells including IL-1,IL-6, and TNF-α. Similarly, fungi and viruses may also contain antigenicmolecules and TLR agonists that may trigger inflammation in a local orsystemic manner, depending on the identity of the microbe, itspathogenic potential, and extent of overgrowth or metabolic activity orin cases of acute a chronic infection or intoxification.

The compositions disclosed herein may comprise one or more chelator andone or more base. Said compositions may also comprise one or morechelator and one or more base and one or more enhancer.

A. Chelators

Chelators may comprise heterogeneous synthetic or naturally-occurringmolecules capable of coordinating, and in some cases, binding cargo.Examples of cargo comprise, without limitation, metals, lipids, volatilearomatic compounds, carbohydrates, microbial components and microbes.Chelators may have more than one donor atom that may coordinate a singlecargo. Chelators may also be able to coordinate more than one cargo andis dependent on chelator size and coordination groups. Chelators may beclassified according to the number of donor atoms correctly positionedfor potential binding to a Lewis acid or other cargo. Chelators may bebidentate, tridentate and polydentate in nature (for example, EDTA ishexadentate). Additionally, chelators may by macrocyclic whereby a largering compound may contain several donor atoms that may bind a Lewis acidinside said ring structure. A chelator:cargo complex is called achelate.

Examples of suitable chelators that may be used in said compositioninclude, but are not limited to, cyclodextrins, α-cyclodextrin,β-cyclodextrin, γ-cyclodextrin, δ-cyclodextrin, methyl-β-cyclodextrin,ethylenediamine, diethylenetriamine, EDTA, EGTA, thiocyanate, porphine,heme, nitrilotriacetic acid, rhodotorulic acid, citrate, anachelins,ferrioxamines, deferoxamine mesylate, cellulose, methylcellulose,carboxymethylcellulose, citrus pectins, apple pectins, orange pectins,carrot pectins, amidated pectins, amidated low methoxyl pectin,high-ester pectins, low-ester pectins, microbial cellulosic material,agar, lignin, curcumin, chlorophylls, resveratrol, alginate, tannins,quercetin, oleuropein, HEDTA, DTPA, EDDHA, succimer, dimercaprol andderivatives thereof. Examples of chelator derivatives include but arenot limited to tosylated, acylated, hydroxylalkylated, alkylated,carboxylated, methylated, hydroxypropylated, hydroxylated,sulfoalkylated, amidated, phosphorylated and sulfonated forms and thelike. Said chelators may be used in free or salt form. Non-limitingexamples of chelators and their use are provided in Table 1.

Suitable chelators may be used in liquid, gel, paste, crème and ointmentformulations at concentrations ranging from about 500 mM to about 0.1 nMper dose, depending on the use. Some compositions may comprise chelatorfrom about 500 mM to about 1 mM per dose. Some compositions may comprisechelator from about 500 μM to about 1 μM per dose. Other compositionsmay comprise chelator from about 10 μM to about 10 nM per dose. Yetother compositions may comprise chelator from about 800 nM to about 0.1nM per dose.

Suitable chelators may be used in solid and powder formulations atamounts ranging from about 1,000 mg to about 50 ng per dose, dependingon the use. Some compositions may comprise chelator from about 1,000 mgto about 50 mg per dose. Some compositions may comprise chelator fromabout 1 mg to about 25 μg per dose. Some compositions may comprisechelator from about 10 μg to about 100 ng per dose. Other compositionsmay comprise chelator from about 1 μg to about 50 ng per dose.

B. Bases

Bases may comprise an agent that donates electrons or hydroxide ions,accepts protons, may behave as Arrhenius bases, Brønsted-Lowry bases andLewis bases. Arrhenius bases increase the hydroxyl ion concentration ina solution. Brønsted-Lowry bases accept protons. Lewis bases donateelectron pairs. Bases may comprise primary, secondary, tertiary orcyclic amines.

Examples of suitable bases that may be used in said composition include,but are not limited to, pyridoxine, pyridoxal-5-phosphate, pyridoxineHCl, vitamin B6, vitamin B12, vitamin K, arginine, polyarginines,lysine, polylysines, uracil, sodium bicarbonate, analine, biogenicamines, methylamine, dimethylamine, trimethylamine, ammonia,propylamines, nitroanalines, haloamines, oximes, sodium hydroxide,potassium hydroxide and derivatives thereof. Said bases may have one ormore hydroxyl group, pyridine, pyridine nitrogen group, amine or otherand may comprise: vitamin base, nucleotide base, basic amino acid, basicsugar, basic lipid, basic mineral, basic salt, basic small molecule,basic peptide or basic enzyme. Said base may be used in free or saltform. Non-limiting examples of bases and their use are provided in Table2.

Suitable bases may be used in liquid, gel, paste, crème and ointmentformulations at concentrations ranging from about 250 mM to about 750 pMper dose, depending on the potency and use. Some compositions maycomprise base from about 250 mM to about 1 mM per dose. Somecompositions may comprise base from about 800 μM to about 5 μM per dose.Other compositions may comprise base from about 10 μM to about 100 nMper dose. Yet other compositions may comprise base from about 120 nM toabout 750 pM per dose.

Suitable bases may be used in solid and powder formulations at amountsranging from about 1,000 mg to about 50 ng per dose, depending on thepotency and use. Some compositions may comprise base from about 1,000 mgto about 50 mg per dose. Some compositions may comprise base from about75 mg to about 5 mg per dose. Some compositions may comprise base fromabout 5 mg to about 100 μg per dose. Other compositions may comprisebase from about 150 μg to about 8 μg per dose. Yet other compositionsmay comprise base from about 7.5 μg to about 50 ng per dose.

C. Enhancers

An enhancer may comprise a synthetic or natural agent that enhances theeffects of chelator or base in said composition. Enhancers may act byactivating said chelator or said base; may protect or preserve saidchelator or said base; may promote synergism between said chelator andsaid base; may act in a supplemental manner to promote synergisticeffects between chelator and base to reduce the total dose of eachactive agent used; or other.

Examples of suitable enhancers that may be used in said compositioninclude, but are not limited to, antacids, ion channel antagonists,proton transporter antagonists, bacteriocins, sugar hydrolyzing enzymes,neutral sugars, sugar alcohols, xylitol, mannitol, sorbitol, Stevia,antimicrobial lipids, iron-binding proteins, plant extracts andderivatives thereof. Other non-limiting examples of suitable enhancersmay include extracts and oils of green tea, mint, peppermint, cinnamon,spearmint, clove, aloe, ginger, lemongrass, avocado, olive, pomegranate,acai, and citrus. Other examples of suitable enhancers may include,without limitation, binding compounds such as polyethylene glycol,polyvinyl pyrrolidone, magnesium stearate and others. Yet other examplesof suitable enhancers may be, without limitation, ascorbic acid,phenylalanine, histatins, statherins, proline-rich proteins, lipase,sialoperoxidase, adhesion-modulating proteins, carbonic anhydrases,amylases, peroxidases, lactoferrin, and mucins. Said enhancers do notinhibit that activity of chelator or base in said composition.

Suitable enhancers may be used in liquid, gel, paste, crème andointmente formulations at concentrations ranging from about 100 mM toabout 50 μM per dose, depending on the potency and use. In someembodiments, suitable enhancer concentrations may range from about 100mM to about 5 mM per dose. In some embodiments, suitable enhancerconcentrations may range from about 5 mM to about 50 μM per dose. Insome embodiments, suitable enhancer concentrations may range from about75 μM to about 500 nM per dose. In other embodiments, suitable enhancerconcentrations may range from about 1 μM to about 10 nM per dose. Yet inother embodiments, suitable enhancer concentrations may range from about25 nM to about 50 pM.

Suitable enhancers may be used in solid and powder formulations atamounts ranging from about 5,000 mg to about 10 ng per dose, dependingon the potency and use. In some embodiments, suitable enhancer amountsmay range from about 5,000 mg to about 150 mg per dose. In someembodiments, suitable enhancer amounts may range from about 200 mg toabout 5 mg per dose. In some embodiments, suitable enhancer amounts mayrange from about 5 mg to about 250 μg per dose. In other embodiments,suitable enhancer amounts may range from about 500 μg to about 10 μg perdose. Yet in other embodiments, suitable enhancer amounts may range fromabout 25 μg to about 500 ng. Yet in other embodiments, suitable enhanceramounts may range from about 750 ng to about 10 ng per dose.

Chelators, bases and enhancers, or their acceptable salts may containone or more asymmetric centers and may thus give rise to enantiomers,diastereomers, and other stereoisomeric forms that may be defined, interms of absolute stereochemistry, as (R)− or (S)− or, as (D)− or (L)−for amino acids. The present invention is meant to include all suchpossible isomers, as well as, their racemic and optically pure forms.Optically active (+) and (−), (R)− and (S)−, or (D)− and (L)− isomersmay be prepared using chiral synthons or chiral reagents, or resolvedusing conventional techniques, such as reverse phase HPLC. When thecompounds described herein contain olefinic double bonds or othercenters of geometric asymmetry, and unless specified otherwise, it isintended that the compounds include both E and Z geometric isomers.Likewise, all tautomeric forms are also intended to be included.

Chelators, bases and enhancers or salts thereof may form a solvateand/or a crystal polymorph, and the present invention contains suchsolvates and crystal polymorphs of various types. A solvate means asolvate of the compound of the present invention or its salt, andexample includes solvate of which solvent is alcohol (e.g., ethanol),hydrate, or the like. Example of hydrate includes monohydrate, dihydrateor the like. A solvate may be coordinated with an arbitrary number ofsolvent molecules (e.g., water molecules). The compounds or saltsthereof may be left in the atmosphere to absorb moisture, and a casewhere adsorbed water is attached or a case where hydrate is formed mayarise. Moreover, the compounds or salts thereof may be recrystallized toform their crystal polymorph.

D. Pharmaceutical compositions

Any of the chelator and base compositions disclosed herein may beformulated into a pharmaceutical and may comprise combinations ofchelator and base as the active ingredients. Pharmaceutical compositionsmay also comprise enhancers.

In some compositions comprising a pharmaceutically acceptable dose ofchelator or base or comprising a pharmaceutically acceptable chelator orbase may require FDA approval as a pharmaceutical for use to treat amicrobe-mediated inflammatory disease.

Pharmaceutical compositions may be formulated with appropriatepharmaceutically acceptable excipients, carriers, diluents or vehicles,and may be formulated into preparations in solid, semi-solid, liquid,gels, pastes, suspension, emulsion, or gaseous forms, and may beformulated into a pharmaceutically acceptable dosage form such as:tablets, capsules, caplets, gums, powders, granules, ointments, crèmes,solutions, suspensions, emulsions, suppositories, injections, inhalants,gels, nanoparticles, microspheres, and aerosols. As such, administrationmay be achieved in various ways, usually by oral or topicaladministration. In pharmaceutical dosage forms, the chelator and basemay be administered in the free form or in the form of theirpharmaceutically acceptable salts, or they may also be used in atime-release formulation, or they may be administered sequentially in adiscrete manner, or they may also be used in combination with otherpharmaceutically active compounds.

The term “pharmaceutical dose” or “pharmaceutical dosage form,” refersto physically discrete units suitable as unitary dosages for humans andother mammals, each unit comprising a predetermined quantity of agentsin an amount calculated sufficient to produce the desired effect inassociation with an acceptable diluent, carrier, or vehicle of aformulation. The specifications for the unit dosage forms may depend onthe particular chelator and base combination employed, the effect to beachieved, the route of administration and the pharmacodynamicsassociated with the mammal.

For demonstrating the synergistic activity of the selected chelator andbase agents to be used in a pharmaceutical composition and forestablishing an appropriate fixed-dose ratio for efficacy againstmicrobe-mediated or microbe-enhanced inflammatory diseases, varyingamounts of the two agents are administered to appropriate animal modelsof inflammatory disease, either at a time of active disease (followingdisease onset) or at an early time point representative of pre-clinicaldisease, and the effect on disease activity or progression is measured.Alternatively, the effects of varying amounts of the two agents may betested on a cellular response mediating inflammation that may beinvolved in the pathogenesis of said disease. Alternatively, the effectsof varying amounts of the two agents in various formulations may betested on a microbial response; the presence, absence or degree ofpathogenic effectors; metabolic processes and/or growth that may beinvolved in the pathogenesis of said disease as a means to determine theappropriate dose and ratio for use as a pharmaceutical composition.

In some pharmaceutical compositions, suitable chelators may comprisecyclodextrins, pectins, modified pectins, ethylenediamine,diethylenetriamine, EDTA, EGTA, thiocyanate, porphine, heme,nitrilotriacetic acid, rhodotorulic acid, citrate, anachelins,ferrioxamines, deferoxamine mesylate, their salts, derivatives or other.

In some pharmaceutical compositions, suitable bases may compriseanalines, biogenic amines, methylamine, dimethylamine, trimethylamine,pyridoxine, pyridoxal-5-phosphate, pyridoxamine, ammonia, sodiumbicarbonate, ammonium salts, propylamines, nitroanalines, haloamines,oximes, sodium hydroxide, potassium hydroxide or other. For oralformulations, the agents may be used alone or in combination withappropriate additives to make tablets, powders, granules, gums,lozenges, rapid melt tablets, capsules, gels, pastes, solutions orsuspensions. Said formulations may further comprise conventionaladditives, such as xylitol, mannitol, sorbitol, stevia or the like; withbinders, such as crystalline cellulose, cellulose derivatives, acacia,starches, gelatins or the like; with disintegrators, such as starches,carboxymethylcellulose or the like; with lubricants, such as talc,magnesium stearate or the like; and if desired, with diluents, bufferingagents, moistening agents, preservatives, and flavoring agents.

Acceptable excipients, such as vehicles, adjuvants, carriers ordiluents, are commercially available. Moreover, acceptable auxiliarysubstances, such as pH-adjusting and buffering agents,tonicity-adjusting agents, stabilizers, wetting agents and the like arecommercially available. Any compound useful in the methods andcompositions of the invention can be provided as an acceptablebase-addition salt. “Acceptable base-addition salt” refers to thosesalts that retain the biological effectiveness and properties of thefree acids, which are not biologically or otherwise undesirable. Thesesalts are prepared by adding an inorganic base or an organic base to thefree acid. Salts derived from inorganic bases include, but are notlimited to, the fluoride, sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Preferred inorganic salts are the ammonium, fluoride, sodium, potassium,calcium, and magnesium salts. Salts derived from organic bases include,but are not limited to, salts of primary, secondary, and tertiaryamines, substituted amines including naturally occurring substitutedamines, cyclic amines and basic ion exchange resins, such asisopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like. Exemplaryorganic bases are isopropylamine, diethylamine, ethanolamine,trimethylamine, dicyclohexylamine, choline, and the like.

E. Nutritional supplement compositions

Any of the chelator and base compositions disclosed herein may beGenerally Recognized as Safe (GRAS) and may be used in amounts at orbelow the FDA recommended daily allowance. Said compositions may be usedas a nutritional supplement to promote health. In some embodiments,compositions may be used to promote health in a localized manner and maybe dependent on the route of administration, dose or absorption kineticsand dynamics.

Said compositions may be formulated with acceptable carriers ordiluents, and may be formulated into preparations in solid, semi-solid,pressed powder, powder, liquid, gel, suspension, emulsion, or gaseousforms, and may be formulated into preparations such as liquids, syrups,concentrates, tablets, capsules, caplets, powders, rapid melts, thinstrips, granules, ointments, crèmes, solutions, suspensions, emulsions,suppositories, injections, inhalants, gels, microspheres, nanoparticles,crystals and aerosols. As such, dose administration may be achieved invarious ways, usually by oral administration. In nutritional supplementdosage forms, the chelator and base may be administered in the form oftheir acceptable salts, or they may also be used in a time-releaseformulation, or they may be used in combination with other nutritionalsupplement compounds.

Nutritional supplement compositions may comprise suitable chelators suchas: cyclodextrins such as α-cyclodextrin, β-cyclodextrin,γ-cyclodextrin, δ-cyclodextrin, methyl-β-cyclodextrin, and theirderivatives; celluloses such as methylcellulose, carboxymethylcellulose,cellulosic material from algae and lignin and their derivatives; pectinsfrom citrus, apple, orange, carrot or other; modified pectins such asamidated pectins, amidated low methoxyl pectins, high-ester pectins,low-ester pectins and other; and various other chelators such as agar,curcumin, chlorophylls, resveratrol, alginate, tannins, quercetin,oleuropein, EDTA and EGTA.

Nutritional supplement compositions may comprise suitable bases such as:vitamin bases such as pyridoxine, pyridoxamine, pyrodoxal-5-phosphate,pyridoxine HCl, vitamin B6, vitamin B12, vitamin K; basic amino acidsand derivatives such as arginine, polyarginines, lysine, polylysines,protamine sulfate; uracil; basic salts such as sodium bicarbonate andothers; sodium hydroxide and potassium hydroxide.

For oral formulations, the agents may be used alone or in combinationwith appropriate additives to make tablets, powders, pressed powders,crystals, granules, capsules, gums, lozenges, rapid melt tablets,capsules, gels, thin strips, pastes, solutions or suspensions. Saidformulations may further comprise conventional additives, such asxylitol, mannitol, sorbitol, stevia or the like; with binders, such ascrystalline cellulose, cellulose derivatives, acacia, starches, gelatinsor the like; with disintegrators, such as starches,carboxymethylcellulose or the like; with lubricants, such as talc,magnesium stearate or the like; and if desired, with diluents, bufferingagents, moistening agents, preservatives, and flavoring agents.

The agents may be encapsulated in gelatin capsules together withinactive ingredients and powdered carriers, such as, but not limited to,glucose, lactose, sucrose, sucralose, mannose, mannitol, xylitol,stevia, aspartame, neotame, acesulfame potassium, starch, cellulose orcellulose derivatives, magnesium stearate, stearic acid, sodiumsaccharin, talcum, magnesium carbonate. Examples of additional inactiveingredients that may be added to provide desirable color, taste,stability, buffering capacity, dispersion or other known desirablefeatures include, without limitation, red iron oxide, silica gel, sodiumlauryl sulfate, titanium dioxide, and edible white ink. Similar diluentsmay be used to make compressed tablets. Both tablets and capsules may bemanufactured as sustained release products to provide for continuousrelease of agents over a period of minutes. Compressed tablets may besugar coated or film coated to mask any unpleasant taste and protect thetablet from the atmosphere, or enteric-coated for selectivedisintegration in the gastrointestinal tract. Liquid dosage forms fororal administration may contain coloring and flavoring to increaseaesthetic experience.

Compressed tablets may be generated using a process known as wetgranulation, whereby the active agents may be mixed in powder form withinactive ingredients. Once mixed, the powder may be forced through amesh screen in a process called dry screening. The mixture may thenmolded into a tablet using mechanical compression by a punch and diesystem. The final tablet may then be coated for aesthetics. Compressedtablets may also be generated using a process using a tablet press,whereby the active agents may be mixed in dry powder form with inactiveingredients. Once mixed, the dry powder may be molded into a tabletusing mechanical compression by a punch and die system.

For oral administration of dietary supplement compositions to themucosal surfaces of the mouth, throat and upper digestive tract, theactive agents with other suitable agents may be made into formulationssuch as liquids, fast melting capsules, fast-dissolving tablets,powders, gels, thin strips and lozenges. In one embodiment, acomposition may be formulated into a fast-dissolving thin strip and maycomprise dehydrated polymer to act as an excipient, such as cellulose,gelatin or starch that, when hydrated under the tongue with saliva maydissolve to release the chelator and base. In another embodiment, acomposition may be formulated into a fast-dissolving lozenge and maycomprise liquid paraffin, sugar-substitutes, sugar alcohols,non-crystallizing sorbitol solution, flavoring agent, coloring agent orthe like, and may release the chelator and base to the tongue, throatand esophagus. In another embodiment, a composition may be formulatedinto a fast-dissolving tablet and may comprise disintegrant, filler,sugar alcohol, flavoring agent, coloring agent or the like, and mayrelease the chelator and base to the tongue, teeth, gums and mucosalsurfaces of the mouth. In another embodiment, a composition may beformulated into a gel and may comprise cellulose, gelatin, pectin orother polymer, sugar alcohols, flavoring agent, coloring agent or thelike and may release the chelator and base to the surfaces of the mouth,teeth, tongue, throat, and esophagus.

Acceptable excipients, such as vehicles, adjuvants, carriers ordiluents, are commercially available. Moreover, acceptable auxiliarysubstances, such as pH-adjusting and buffering agents,tonicity-adjusting agents, stabilizers, wetting agents and the like arecommercially available. Any compound useful in the methods andcompositions of the invention can be provided as an acceptablebase-addition salt. “Acceptable base-addition salt” refers to thosesalts that retain the biological effectiveness and properties of thefree acids, which are not biologically or otherwise undesirable. Thesesalts are prepared by adding an inorganic base or an organic base to thefree acid. Salts derived from inorganic bases include, but are notlimited to, the fluoride, sodium, potassium, lithium, ammonium, calcium,magnesium, iron, zinc, copper, manganese, aluminum salts and the like.Preferred inorganic salts are the ammonium, fluoride, sodium, potassium,calcium, and magnesium salts. Salts derived from organic bases include,but are not limited to, salts of primary, secondary, and tertiaryamines, substituted amines including naturally occurring substitutedamines, cyclic amines and basic ion exchange resins, such asisopropylamine, trimethylamine, diethylamine, triethylamine,tripropylamine, ethanolamine, 2-dimethylaminoethanol,2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine,caffeine, procaine, hydrabamine, choline, betaine, ethylenediamine,glucosamine, methylglucamine, theobromine, purines, piperazine,piperidine, N-ethylpiperidine, polyamine resins and the like. Exemplaryorganic bases are isopropylamine, diethylamine, ethanolamine,trimethylamine, dicyclohexylamine, choline, and caffeine.

F. Food Safety, Cosmetic, and Surface-Cleaning Compositions

Any of the chelator and base compositions disclosed herein may be GRASagents and may be used as food additives to promote health. Foodadditive compositions may be formulated may be formulated as foodsprays, food washes or concentrates. Food safety formulations may beused to promote health or to limit undesired microbial metabolism andthus may serve as food safety agents, food preparatory agents and/orfood preservatives.

Some food safety compositions may be used directly on food surfaces ormay be used as preservative in food products. Said compositions maycomprise EDTA, sodium pyrophosphate and gamma-cyclodextrin as chelator;and pyridoxal-5-phosphate, NaOH, lysine and sodium pyrophosphate asbase. Compositions may comprise chelator concentration at about 0.1 mMto about 0.1 μM and base concentration at about 0.1 mM to about 0.1 μM.

Additionally, food safety formulations may be applied frequently or onan as needed basis to surfaces or objects such as: cutting board,kitchen counter, kitchen sink, food-processing devices such as conveyerbelts, knives, forks, hooks, gloves or other and food storage devicessuch as packaging, containers, boxes or other.

In some embodiments, food safety compositions may comprise EDTA, EGTA,rhodotorulic acid and ferrichrome as chelator; protamine sulfate, trisamine, tris (hydroxymethyl) aminomethane hydrochloride, diethylaminehydrochloride, KOH and NaOH as base; and boric acid, and sodiumorthovanadate as enhancer. In some surface care compositions acceptablechelator concentrations may range from about 100 mM to about 1 μM,acceptable base concentrations may range from about 500 μM to about 1 μMand acceptable enhancer concentrations may range from about 50 μM toabout 10 nM.

In other food safety compositions for use on food preparation surfacesand food storage surfaces may comprise ferrichrome, EDTA andrhodotorulic acid as chelator; KOH, NaOH and sodium bicarbonate as baseand sorbic acid as enhancer. In said compositions, EDTA may be used atconcentrations of ranging from about 0.1 M to about 0.1 mM, rhodotorulicacid may be used at concentrations of about 10 μM to about 10 nM,ferrichrome may be used at concentrations of about 5 mM to 5 μM; KOH andNaOH may be used at concentrations of about 0.8 M-0.5 μM; sodiumbicarbonate may be used at about 0.1 M to about 0.1 mM; sorbic acid maybe used at concentrations of about 0.5 mM to about 0.5 μM.

Any of the chelator and base compositions disclosed herein may be GRASagents and may be suitable for formulation in a cosmetic to limitundesired microbial growth, undesired microbial metabolism and thusserve as a preservative and/or a deodorant. Cosmetic compositions may beformulated in an elution patch, wipe, pad, sponge, cloth, strip, gel,paste, solution, emollient, serum, moisturizer, make-up, lotion,shampoo, conditioner, gel, mousse, soap, deodorant, antiperspirant,spray, wash and solution and may be used for topical administration.

In some compositions, acceptable chelator and base may be used forveterinary applications such as in formulations for promoting oralhealth, wound care or other. Veterinary formulations may include sprays,gels, liquids, syrups, tablets, capsules, lotions, salves, or other.

Any of the chelator and base compositions disclosed herein may be usedas surface cleaners, disinfectants, or other. Surface treatments may beformulated as concentrates, solutions, sprays, powders, wipes or other.Surfaces may include: biologic surfaces comprising oral, gut, nasal,ocular, otic, topical, food or other; inanimate surfaces comprisingstructural surfaces, clothing, gloves, diapers, undergarments, kitchensurfaces, bathroom surfaces, painted surfaces, tiled surfaces, doorknobs, handles, computers, remote controls, steering wheels, cellphones, manufacturing machinery, hospital surfaces, devices or other.

G. Health-promoting compositions

Associations between chronic oral infections and oral inflammation andrheumatologic disease, autoimmune diseases neurodegenerative diseases,respiratory diseases, cardiovascular diseases and stroke, andlow-birth-weight/premature births are being realized (U.S. SurgeonGeneral, WHO). Microbe-mediated and microbe-enhanced inflammatorydiseases may include autoimmune diseases including rheumatoid arthritis,Crohn's disease, psoriasis, psoriatic arthritis, systemic lupuserythematosus, oral lichen planus, Behçet's disease, Sjogren's syndromeand other autoimmune diseases; degenerative diseases includingosteoarthritis, stroke, cardiovascular disease, atherosclerosis,Alzheimer's disease, other dementias, macular degeneration, chronicobstructive pulmonary disease, halitosis, irritable bowel disease andother degenerative diseases; chronic infections including humanimmunodeficiency virus infection, hepatitis C virus infection,cytomegalovirus infection, otitis media, periodontal disease, cavities,sinusitis, rhinitis, pneumonia, rheumatic fever, endocarditis,pericarditis, and other viral, bacterial, fungal, parasite and otherinfection. Accordingly, any of the chelator and base compositionsdisclosed herein may be used to promote one or more of oral health,cardiovascular health, mental health, and/or systemic health.

Oral Health.

Oral health relates to the balance of beneficial and pathogenicactivities in the oral and nasopharyngeal cavities and is criticallydependent on the relationship between the microbial flora and mucosalimmunity. Diseases such as, but not limited to, cavities (CV),gingivitis (GV), periodontal disease (PD), rheumatic fever (RF), otitismedia (OM), sinus infection (SI), chronic rhinosinusitis (CRS), chronichalitosis (HT) and pneumonia (PN) may stem from oral imbalance, oralmicrobial overgrowth, microbial effectors, undesirable microbialmetabolic products and/or inflammation.

Cardiovascular Health.

Cardiovascular health relates to the balance of beneficial andpathogenic activities in the cardio-pulmonary-vascular system. Build-upof inflammatory plaques and/or chronic infections of the heart,cardio-pulmonary and/or vasculature may arise from microbial biofilmformation, microbial infection, microbial effectors, undesirablemicrobial metabolic products and/or inflammation. Non-limiting examplesof cardiovascular diseases with microbial and microbial-relatedinflammation may include: Rheumatic fever (RF), atherosclerosis (AS),stroke (ST), cardiovascular disease (CVD) and chronic obstructivepulmonary disease (COPD).

Mental Health.

Mental health relates to the balance between beneficial and pathogenicactivities of the brain and central nervous system and may be directlyor indirectly impacted by build-up of inflammatory plaques and/orchronic infections of the brain and central nervous system and may arisefrom microbial biofilm formation, microbial infection, microbialeffectors, undesirable microbial metabolic products and/or inflammation.Non-limiting examples of mental health conditions comprise Alzheimer'sdisease (AD), dementias (DM), depression, Parkinson's disease,amyotrophic lateral sclerosis and others.

Systemic Health.

Systemic health relates to the balance of beneficial and pathogenicactivities of organs, organ systems and tissues of the body and may bedirectly or indirectly impacted by build-up of microbial overgrowth,inflammatory plaques and/or chronic infections in the body. Non-limitingexamples of systemic health conditions comprise Behcet's Disease (BD),Sjogren's Syndrome (SS), lichen planus (LP), systemic lupuserythematosus (SLE), Rheumatoid arthritis (RA), and other autoimmunediseases having disease manifestations involving microbes ormicrobe-mediated inflammation; degenerative diseases such as irritablebowel disease (IBD), colitis (CO), osteoarthritis (OA), diabetes,metabolic diseases and cancers.

Examples of chelator and base compositions for use in promoting healthmay comprise one or more chelator, base and enhancer. Compositions maycomprise EGTA, EDTA, citrate, cyclodextrins, vitamin B12, pectins anddeferoxamine mesylate as chelator; KOH, NaOH, lysine, arginine, sodiumbicarbonate, α-galactase, pyridoxal-5-phosphate, pyridoxine,pyridoxamine, vitamin B6, lysozyme and vitamin K as base; and ascorbicacid, proline, uracil, remitidine, famotidine, omeprazole and other acidblockers, plant extracts and zinc chloride as enhancer. Commerciallyavailable flavoring agents, coloring agents, fillers, excipients,carriers, and buffers may be formulated into health-promotingcompositions.

III. Methods of the Invention

A. Methods for Promoting Oral Health

Any of the chelator and base compositions (e.g., pharmaceuticalcompositions) disclosed herein can be used in methods to promote oralhealth. Many microbial metabolic products have beneficial effects formammals (examples are fermented foods and beverages). Many othermicrobial metabolic products have pathogenic effects on mammals (Ramseyet al. (2011) PLOS Pathogens). In some embodiments, administration ofcompositions comprising chelator and base can affect microbialmetabolism, namely acid production. Non-limiting examples of oralpathologies capable of treatment by the methods disclosed hereinemploying any of the compositions disclosed herein include cavities(CV), gingivitis (GV), periodontal disease (PD), rheumatic fever (RF),otitis media (OM), sinus infection (SI), chronic rhinosinusitis (CRS),chronic halitosis (HT) or pneumonia (PN).

In some aspects, said method can comprise: (i) identifying an individualat increased risk for developing or exhibiting signs of oral disease,(ii) measuring pH, leukocyte esterase, nitrate, microbial marker levels,or plaque-staining dye retention in the mouth of the individual, (iii)comparing said levels to healthy levels to determine if oral pathologyis present, and (iv) if evidence of oral pathology is present,contacting an effective amount of a composition comprising a chelatorand a base to the oral surface.

In another aspect, any of the chelator and base compositions (e.g.,pharmaceutical compositions) disclosed herein may be used in methods forlimiting or eliminating microbes and/or microbial biofilms in an oralcavity of an individual comprising contacting a surface in the oralcavity with an effective amount of any of the compositions disclosedherein.

B. Methods for Promoting Cardiovascular Health

Any of the chelator and base compositions (e.g., pharmaceuticalcompositions) disclosed herein can be used in methods to promotecardiovascular health. Non-limiting examples of cardiovascularpathologies capable of treatment by the methods disclosed hereinemploying any of the compositions disclosed herein include Rheumaticfever (RF), atherosclerosis (AS), stroke (ST), cardiovascular disease(CVD) and chronic obstructive pulmonary disease (COPD). In some aspects,said method can comprise: (i) identifying an individual at increasedrisk for developing or exhibiting signs of a cardiovascular disease and(ii) administering a clinically effective amount of any of thecompositions disclosed herein to the individual. In other aspects, saidmethod comprises administering a clinically effective amount of any ofthe compositions disclosed herein to an individual diagnosed with orthought to be afflicted with a cardiovascular disease.

C. Methods for Promoting Mental Health

Any of the chelator and base compositions (e.g., pharmaceuticalcompositions) disclosed herein can be used in methods to promote mentalhealth. Non-limiting examples of mental or neurological pathologiescapable of treatment by the methods disclosed herein employing any ofthe compositions disclosed herein include Alzheimer's disease (AD),dementias (DM), depression, Parkinson's disease, and amyotrophic lateralsclerosis. In some aspects, said method can comprise: (i) identifying anindividual at increased risk for developing or exhibiting signs of amental disease and (ii) administering a clinically effective amount ofany of the compositions disclosed herein to the individual. In otheraspects, said method comprises administering a clinically effectiveamount of any of the compositions disclosed herein to an individualdiagnosed with or thought to be afflicted with a mental or neurologicaldisease.

D. Methods for Promoting Systemic Health

Any of the chelator and base compositions (e.g., pharmaceuticalcompositions) disclosed herein can be used in methods to promotesystemic health. Non-limiting examples of systemic pathologies capableof treatment by the methods disclosed herein employing any of thecompositions disclosed herein include Behcet's Disease (BD), Sjogren'sSyndrome (SS), lichen planus (LP), systemic lupus erythematosus (SLE),Rheumatoid arthritis (RA), and other autoimmune diseases having diseasemanifestations involving microbes or microbe-mediated inflammation;degenerative diseases such as irritable bowel disease (IBD), colitis(CO), osteoarthritis (OA), diabetes, metabolic diseases and cancers. Insome aspects, said method can comprise: (i) identifying an individual atincreased risk for developing or exhibiting signs of a systemic diseaseand (ii) administering a clinically effective amount of any of thecompositions disclosed herein to the individual. In other aspects, saidmethod comprises administering a clinically effective amount of any ofthe compositions disclosed herein to an individual diagnosed with orthought to be afflicted with a systemic disease (such as, but notlimited to an autoimmune disease or an inflammatory disease).

E. Administration

Any of the chelator and base compositions (e.g., pharmaceuticalcompositions) disclosed herein may be administered on a frequent basisto affect microbial growth, microbial metabolism, microbial biofilmintegrity, microbial biofilm production, microbial toxin production ormicrobial acid production. Said compositions may be administered on afrequent basis to affect microbial effectors and thus reduceinflammation. Said compositions may be administered on a frequent basisfor the promotion and maintenance of health.

It is within the level of skill of a clinician or medical researcher orveterinarian to determine the preferred route of administration and thecorresponding dosage form and amount, as well as the dosing regimen,i.e., the frequency of dosing. In some embodiments, the composition maybe delivered in multi-dosing format whereby said composition may beadministered several times a week, once a day, twice a day, three timesa day, or more to achieve the appropriate therapeutic level. However,this generalization does not take into account such important variablesas the specific type of microbial species to be affected, the specificinflammatory disease, the specific therapeutic agent involved and itspharmacokinetic profile, and the specific individual involved. For otherapproved products in the marketplace, much of this information isalready provided by the results of clinical studies carried out toobtain such approval. In other cases, such information may be obtainedin a straightforward manner in accordance with the teachings andguidelines contained in the instant specification taken in light of theknowledge and skill of the artisan. The results that are obtained mayalso be correlated with data from corresponding evaluations of past andcurrent marketed product(s) utilizing comparable clinical testingmethods.

Frequency of administration may be once a month, once a week, once aday, up to 3 times per day, up to 10 times a day, before bed and on anas-needed basis. Frequency of administration may be dependent on theidentity and concentration of the base and chelator in said composition;and may be dependent on disease risk assessment, disease severity, testresults, or personal preference; and may be dependent on formulation.

Individuals and other mammal at increased risk for development of amicrobe-mediated or microbe-enhanced inflammatory disease, withearly-stage of disease, or with established disease may be treated witha clinically effective amount of any of the compositions disclosedherein to prevent the development of disease, to prevent the progressionof disease, and to prevent the progression of the symptoms or signs ofdisease. The total of a dose of base may generally range from about0.00003 to about 5 mg/dose and the total of a multi-day dose may rangebetween about 0.0003 to about 25 mg/day. The total of a dose of chelatormay generally range from about 0.0005 to about 10 mg/dose and the totalof a multi-day dose may range between about 0.003 to about 30 mg/day.The total dose of enhancer may vary and may generally range from about0.00001 to 15 mg/dose.

In some embodiments, an individual may be administered any of thehealth-promoting compositions disclosed herein, wherein saidcompositions can comprise a chelator agent of at least about 0.003 mg,at least about 0.006 mg, at least about 0.01 mg, at least about 0.03 mg,at least about 0.06 mg, at least about 0.1 mg, at least about 0.125 mg,at least about 0.25 mg, at least about 0.5 mg, at least about 0.75 mg,at least about 1.0 mg, at least about 3.0 mg, at least about 5.0 mg, atleast about 10.0 mg, at least about 25 mg, at least about 50 mg, notmore than about 5.0 g; and base agent in a single dose of at least0.0003 mg, at least about 0.0006 mg, at least about 0.001 mg, at leastabout 0.003 mg, at least about 0.006 mg, at least about 0.01 mg, atleast about 0.03 mg, at least about 0.06 mg, at least about 0.1 mg, atleast about 0.3 mg, at least about 0.6 mg, not more than about 30.0 mg.

In some embodiments, an individual may be administered any of thehealth-promoting compositions disclosed herein, wherein saidcompositions can comprise cyclodextrin or a cyclodextrin derivative in asingle dose of at least about 0.003 mg, at least about 0.006 mg, atleast about 0.01 mg, at least about 0.03 mg, at least about 0.06 mg, atleast about 0.1 mg, at least about 0.125 mg, at least about 0.25 mg, atleast about 0.5 mg, at least about 0.75 mg, at least about 1.0 mg, atleast about 3.0 mg, at least about 5.0 mg, at least about 10.0 mg, atleast about 25 mg, at least about 50 mg, not more than about 5.0 g; andvitamin B6, pyridoxal-5-phosphate, pyridoxamine, pyridoxine or anothervitamin base in a single dose of at least 0.0003 mg, at least about0.0006 mg, at least about 0.001 mg, at least about 0.003 mg, at leastabout 0.006 mg, at least about 0.01 mg, at least about 0.03 mg, at leastabout 0.06 mg, at least about 0.1 mg, at least about 0.3 mg, at leastabout 0.6 mg, not more than about 30.0 mg; and sodium bicarbonate orother basic salt in a single dose of at least 0.0003 mg, at least about0.0006 mg, at least about 0.001 mg, at least about 0.003 mg, at leastabout 0.01 mg, at least about 0.03 mg, at least about 0.06 mg, at leastabout 1.0 mg, at least about 5.0 mg, at least about 25 mg, at leastabout 50 mg, at least about 100 mg, at least about 250 mg, at leastabout 800 mg, at least about 1000 mg, at least about 2000 mg, not morethan about 8.0 g.

Any of the chelator and base compositions (e.g., pharmaceuticalcompositions) disclosed herein may be administered in a manner todeliver base and chelator agents in combination or in sequence. In someembodiments, health-promoting compositions may be administered in aperiodic manner. Administration of compositions may occur at least oncedaily, weekly or monthly or on an as needed basis. Health-promotingcompositions may comprise pharmaceutically acceptable formulations (suchas carriers or excipients) or may be nutritional supplementformulations. Health-promoting compositions may be commercialized inindividual dose forms or in multi-dosing forms.

Methods of use for food safety formulation comprise of contacting thecomposition to a surface prior to, periodically during and/orimmediately following the preparation of food and food products.Preferred methods involve contacting said surface with said compositionsfor a minimum of 5 seconds. Preferred methods also involve the periodicrinsing of contacted surfaces to effectively remove the neutralizedmicrobes, biofilms and metabolic products.

F. Methods for Selecting Base and Chelator Compositions

Methods of selecting base and chelator compositions depend on use.Compositions may be determined by ranking base, chelator and enhanceragents, from most desired to least desired, for multiple parameters suchas safety, solubility, compatibility, pH, potency, taste, consistency,smell and others. Examples of ranking base and chelator agents fordetermining selection in a composition for a desired use are presentedin Tables 1 and 2.

Non-toxic chelator and base compositions may be ranked according to thedesired parameters of potency and approved-for-oral-consumption. Adesired parameter such as potency for a candidate chelator or base maybe given a score of “1” for most desired and a score of “3” for leastdesired. In one embodiment, candidate chelators, bases and enhancers maybe ranked by FDA approval for oral consumption. Agents with a score of“1” may have approval for the desired indication and a score of “3” maynot have approval or may not be approved for the desired indication. Theresultant overall score may be tabulated and may be used to facilitateselection. In this example, the lowest overall score represents the bestcandidates for use in a non-toxic composition.

TABLE 1 CHELATORS THAT MAY BE USED IN COMBINATION WITH BASE AGENTSANTICIPATED CHELATOR POTENCY{circumflex over ( )} CONSUMABLE USE*Resveratrol 2 1 O, T, V, F, S Quercetin 1 1 O, T, V, F, S Curcumin 2 1O, T, V, F, S Oleuropein 2 1 O, T, V, F, S Inositol 2 1 O, T, V, F, Shexaphosphate EGCG 1 1 O, T, V, F, S Amidated Pectins 1 1 O, T, V, F, SAlginate 1 1 O, T, V, F, S Tannic acid 1 1 O, T, V, F Chitosan 2 2 O, T,V, F, S Exopolysaccharide 1 2 F, S Cellulosic material 1 2 O, T, V, F, SPectins 1 1 O, T, V, F, S Citrate, Citrate salts 3 1 O, T, V, F, SAnachelins 1 3 S Ferrioxamines 1 2 O, T, V, F, S Rhodotorulic acid 1 2O, T, V, F, S Microbial cellulose 1 2 O, T, V, F, S Celluloses 2 1 O, T,V, F, S Deferoxamines 1 1 O, T, V, F, S Deferasiroxamine B 1 1 O, T, V,F, S EDTA 1 1 O, T, V, F, S EGTA 1 2 T, V, F, S Chlorophylls 1 2 O, T,V, F, S DMSA 1 2 T, V, F, S DMPS 1 2 T, V, F, S Vitamin B12 1 1 O, T, V,F, S Cyclodextrins 1 1 O, T, V, F, S {circumflex over ( )}Score 1-3. 1,most; 2, moderate; 3, least. *O: oral administration, T: topicaladministration, V: veterinary, F: food safety, S: surfaces.

TABLE 2 BASE AGENTS THAT MAY BE USED IN COMBINATION WITH CHELATORSANTICIPATED BASE POTENCY{circumflex over ( )} CONSUMABLE USE* Pyridoxine1 1 O, T, V, F, S Pyridoxal-5- 1 1 O, T, V, F, S phosphate Vitamin K 1 1O, T, V, F, S Lysine 2 1 O, T, V, F, S Arginine 2 1 O, T, V, F, S Uracil3 1 O, T, V, F, S Lysozyme 1 2 O, T, V, F, S Tris amine 2 3 T, V, F, STris 2 3 T, V, F, S (hydroxymethyl) amine Sodium bicarbonate 3 1 O, T,V, F, S Protamine sulfate 2 3 O, T, V, F, S D-maltose 1 1 O, T, V, F, SA-galactase 1 1 O, T, V, F, S Remitidine 1 1 O, T, V, F, S Omeprazole 11 O, T, V, F, S Famotidine 1 1 O, T, V, F, S Diethylamine 2 3 F, SSodium phosphate 3 1 O, T, V, F, S forms Dibasic calcium 3 1 O, T, V, F,S phosphate Vitamin B12 2 1 O, T, V, F, S Sodium hydroxide 1 2 O, T, V,F, S Potassium 1 2 O, T, V, F, S hydroxide Pyridoxamine 1 1 O, T, V, F,S {circumflex over ( )}Score 1-3. 1, most; 2, moderate; 3, least. *O:oral administration, T: topical administration, V: veterinary, F: foodsafety, S: surfaces.

Additionally, methods of selecting base, chelator and enhancers for acomposition may be determined experimentally by administering aconcentration or concentration range of agents to microbes, contaminatedsurfaces, animal models and/or mammalian cells to achieve a desiredoutcome. In some embodiments, compositions may be administered to acommensal microbe such as Lactobacillus acidophilus, a pathogenicmicrobe such as Streptococcus mutans, and human biofilms and may beassessed for efficacy. In some embodiments, compositions may beadministered to a mammal or to mammalian cells and the effect of acomposition may be evaluated for efficacy. In some applications,compositions may be selected for having optimal anti-pathogen effectswith minimal affect to commensal microbes. In some applications,compositions may be selected for having optimal anti-biofilm efficacy.In some applications, compositions may be selected for having optimalanti-growth efficacy. In other applications, compositions may beselected for having optimal effects on microbial metabolism. Yet inother applications, compositions may be selected for having optimalactivity against microbe-mediated inflammation.

Additionally, methods for selecting appropriate chelator and basecombinations may be achieved through the use of a test. A test may beconducted to identify and quantify parameters such as: pH; sugarcontent; protein content; the presence of pathogens and/or pathogenicmolecules; the presence and/or level of metabolites such as nitrites,nitrates, sulfates, volatile aromatic compounds; the presence and/orlevel of inflammatory mediators; and others. Methods may relate toconducting a test to identify an optimal composition for use in aspecific indication. Methods may also relate to conducting a test todetermine the frequency in which composition may be used for optimalresults. Methods may also relate to testing frequency to monitor theefficacy of a selected composition and its efficacy over time. Methodsmay relate to sampling a patient, mammal, or surface followed byperforming one or more test on a sample. Methods also relate toperforming one or more test in situ through the administration of a testarticle to a patient, mammal, or surface.

In some embodiments, any of the methods disclosed herein can compriseuse of a pH test strip to measure the pH of a sample. Samples maycomprise environmental, surfaces, saliva, crevicular fluid, urine,sweat, tears, sebaceous secretions, blood or other. In one embodiment, atest may be conducted using a pH test strip to measure the pH of asample to help guide the selection of an optimal base and chelatorcomposition. In one embodiment, a test may be conducted using a pH teststrip to measure the pH of a sample to help guide the dose, frequency ofadministration or other of a composition. In one embodiment, a test maybe conducted using a pH test strip to measure the pH of a sample tomeasure the change in pH following administration of a composition andthus may be used to test the efficacy of a composition.

In some embodiments, any of the methods disclosed herein can compriseuse of stains to test for the presence or burden of microbes, microbialbiofilm, microbial metabolite, microbial effector or inflammation. Insome embodiments, a test may be conducted using a dextran-staining dyeto measure the presence and abundance of biofilm in a sample or on asurface; may be used to help guide the selection of an optimal base andchelator composition; may be used to assess burden and may be to helpguide the dose of composition to be administered; the frequency ofadministration or may be used to measure the change in biofilm burdenfollowing administration of a composition and thus may be used to testthe efficacy of a composition.

Other non-limiting examples of tests relate to the use of immunoassaysto detect and quantify the amount of a specific analyte in a sample. Insome embodiments, immunoassay tests may be approved for use in thediagnosis of a microbial-related inflammatory disease. Examples ofanalytes to be measured may comprise: pathogen markers, metabolicmarkers, microbial effectors, toxins and inflammatory markers such asinflammatory cytokines, chemokines, antigen-specific IgG, IgA, IgM andIgE, and leukocyte esterase and other immune-related enzymes. In someembodiments, test results may be used to help guide the selection of anoptimal base and chelator composition; may be used to assess diseaseseverity and may be to help guide the dose of composition to beadministered; the frequency of administration or may be used to measurethe change in disease status following administration of a compositionand thus may be used to test the efficacy of a composition.

Yet other non-limiting examples of tests relate to the use of chemicaltests to detect and quantify the amount of specific analyte in a sample.In some embodiments, chemical tests may be approved for use in thediagnosis of a microbial-related inflammatory disease. Examples ofanalytes to be measured comprise glucose, nitrite, nitrate, sulfite,sulfate, aromatic volatile compounds or other. In some embodiments, testresults may be used to help guide the selection of an optimal base andchelator composition; may be used to assess disease severity and may beto help guide the dose of composition to be administered; the frequencyof administration or may be used to measure the change in disease statusfollowing administration of a composition and thus may be used to testthe efficacy of a composition.

EXAMPLES Example 1. Compositions to Affect Microbial Growth

Compositions comprising chelator and base can affect microbial growth.It was discovered that the administration of compositions comprisingchelator and base inhibit pathogen growth in a biofilm. Static culturesof pathogen S. mutans, commensal L. acidophilus (purchased from ATCC) orcomplex biofilms isolated from human dental plaque by sterile swab andwere grown for 24-72 h in polystyrene microtiter plates and allowed toform biofilms. Wells were treated briefly with compositions or controlsprior to repletion with sterile media (RPMI, pH 7.2) and their effectson pathogen growth was then evaluated using standard microbiologicalmethods for plating and counting colony forming units (CFUs) on TSAplates (FIGS. 1, 5-6). Optimal microbial growth-interruptingcompositions were identified for each pathogen and for the microbesresiding in the donor plaque by screening chelator, base and enhanceragents at various concentrations and combinations.

An amino acid may be used in combination with a chelator to affectmicrobial growth. The more basic a side chain is (at neutral pH) on anamino acid, the more potential growth affecting potential exists whenused in combination with a chelator (FIG. 6). A chelator may be, forexample, a siderophore such as rhodotorulic acid, rhizoferrin,azotobactin, ferrichrome, desferrioxamine B, bacillibactin, and others.In some embodiments, a chelator may be plant-derived such as pectin,carboxymethylcellulose, cellulose, cyclodextrin and others.

Furthermore, it was discovered that the administration of compositionscomprising chelator and base can inhibit the growth of pathogens inestablished biofilms. Briefly, microbial biofilm was harvested fromhuman mucosal and dermal surfaces and cultured in 96-well flat-bottompolystyrene microtiter plates and spiked with ˜103 CFU of pathogen.Pathogen-containing biofilms were briefly exposed to saline control orcompositions and were then repleated with RPMI media, pH 7.2. Pathogengrowth in saline-treated biofilms was compared to pathogen growth incomposition-treated biofilms. Pathogens were recovered from treatmentsamples by serial dilution in sterile saline followed by plating on TSAblood agar plates. Recovered CFUs were tabulated. Exemplary data arepresented in FIG. 6 and demonstrate that the application of chelator andbase compositions can be effective at limiting the growth of pathogens,even in the presence of a protective biofilm.

Example 2. Compositions to Affect Biofilms

Compositions comprising chelator and base can affect biofilm formation,development, composition and/or integrity. Microbial biofilms are linkedto persistent infection, chronic inflammation and chronic diseases.Clearance or containment of microbial biofilms is important and thereare limited compounds capable of killing pathogens that reside inbiofilms due to poor penetration and/or slow kinetics of antibioticsinto biofilms, enabling pathogen survival and disease persistencedespite the presence of many antibiotics and antiseptics on the market(Brown et al. (1988) J Antimicrob Chemother. 22:777-780; Lewis K. (2001)Antimicrob Agents Chemother. 45:999-1007).

Microbes were cultured in growth media and plated at 10³ CFU/well in a96 well microtiter plate and cultured for 24 h. Wells were then treatedwith 10-50 μL of media only as control or with media+chelator agents,base agents, acid agents, neutral agents or enhancer agents for 30 minat RT followed by repleation in 100 μL of fresh RPMI pH 7.2. Plates werethen incubated for an additional 48-72 h prior to harvest. Wells werewashed 3× in PBS and residual biofilm was stained using crystal violet.Wells were washed 4× in PBS and the amount of retained stain wasmeasured. It was initially determined that strong base alone may inhibitbiofilms, however, strong base is toxic to most organisms and suchcaustic compounds are not meant for frequent exposure. Unexpectedly, itwas observed that biofilms were inhibited by the administration of achelator at pH of 7.0-8.0 (human physiologic pH), but not below pH 6.8.Furthermore, it was determined that low concentrations of base (0.8-1.6μM) in the presence of low concentrations of chelator (1 μM) is superiorto higher concentrations of base (50 μM) or higher concentrations ofchelator (100 μM) alone in blocking biofilms. Synergistic effects wereevaluated and numerous compounds were examined for their ability tosynergistically act with chelators to inhibit biofilm formation(examples of some test compounds analyzed for synergy with chelators arepresented in Table 2). An example of screening results is presented inFIG. 2. Additional dose titrations studies were performed for selectcompounds (examples of dose titration are presented in FIG. 3). It wasdiscovered that compositions comprising low concentrations of chelatorand base, or compositions comprising low concentrations of chelator,base and enhancer, were significantly more effective than higher dosesof the individual agents (FIGS. 1-3). These data present some examplesof chelator and base compositions having biofilm affecting activity.These data also present some examples of chelator and base compositionsthat may be used to inhibit biofilm formation, development, compositionand/or integrity.

Example 3. Compositions to Affect Microbial Metabolism

Compositions comprising chelator and base can affect microbialmetabolism. Many microbes undergo metabolic processes that generateacidic waste products, many of which are secreted into the surroundingenvironment. Sugar fermentation is a metabolic process wherebycarbohydrates are converted into lactic acid, acetic acid or otheracids. Examples where microbial acid production and its secretion intothe surrounding environment have undesired consequences aredemineralization of tooth enamel, metabolic disruption of neighboringmammalian cells (acid erosion of the gums, skin flaking on the scalp(also known as dandruff)), immune cell activation, cancer and others.

It was initially determined that biofilm production can be disrupted bythe administration of a chelator under basic conditions, but not underacidic conditions (FIG. 2). Using similar microtiter plate basedapproach, oral commensals and oral pathogens were cultured in RPMI pH7.2 and were briefly exposed to single base or chelator agents orcombinations of bases and chelators. Assessment of metabolic output 24,48 and 72 hr post-treatment was measured. pH was determined using theacid-sensitive dye Phenol Red to measure acid production in each well,with secondary testing using a pH test strip (pHion and Seimens).Microbial metabolism was also assessed by Multistix 10 SG strips forglucose, nitrite, ketones, protein and other parameters (Siemens). Itwas also determined that low concentration of chelator can inhibit acidproduction by the cariogenic pathogen S. mutans in the presence of base,which significantly increases as increasing base concentrations arepresent (data not shown). It was also discovered that compositions ofchelator and base can disrupt the ability of S. mutans to maintainhomeostatic pH function, as the optimal pH for S. mutans growth andbiofilm formation was determined to be pH 6.5, which is also near theaverage pH of human saliva. For example, at pH input 6.5, the endpointpH was observed to be 8.5; at pH input 8, the endpoint pH was observedto be 8.5 (data not shown).

Furthermore, it was determined that low concentrations of base may alterthe endpoint pH of S. mutans cultures. Screens of base components to usein compositions comprising chelator were performed. Examples of basecomponents screened are presented in Table 2 and FIG. 3. Briefly, the pHof S. mutans cultures were measured 24 and 48 hr post administration ofchelator and candidate base components. Combinations of chelator, baseand enhancer agents were also evaluated for affecting metabolic activityand/or metabolism of S. mutans (FIG. 3). It was also determined that lowconcentrations of base (1×) in the presence of low concentrations ofchelator (1×) were superior to higher concentrations (2×) of base orchelator alone (data not shown). Synergistic effects on microbial acidproduction were observed. These data teach that chelator and base may beadministered to biologic and inanimate surfaces to limit acid productionof the microbes inhabiting them.

Example 4. Compositions and Formulations to Promote Oral Health

Compositions comprising chelator and base can promote oral health. Manymicrobial metabolic products have beneficial effects for mammals(examples are fermented foods and beverages). Many other microbialmetabolic products have pathogenic effects on mammals (Ramsey et al.(2011) PLOS Pathogens). It was observed that S. mutans-specific dextranproduction was enhanced in the presence of chelator at pH 5.5 to 6.5(data not shown). It is also observed that S. mutans-specific dextranproduction can be inhibited when a chelator is administered with a base(FIG. 2). It is also observed that the administration of compositionscomprising chelator and base can affect microbial metabolism, namelyacid production (FIG. 3). Synergistic blockade of microbial dextranproduction and acid production may be important for the prevention oforal pathology such as gingivitis, gum erosion, enamel loss, cavities,periodontitis, mucosal inflammation and others.

In some examples, chelator and base compositions can be formulated in aliquid or solid form and can be administered to mammals to inhibitmicrobial effectors such as acid, biofilm production, overgrowth andrelated inflammation. An example of results obtained from oraladministration of liquid composition (from about 1 to 3 mL) comprisingwater, chelator (gamma-cyclodextrin, 10 μg/mL), base (sodium hydroxide,10 ng/mL) and flavoring agent (cinnamon extract) to human volunteers ispresented in FIG. 4A. Subjects were sampled at intervals over about a12-hour period for salivary pH, protein content and glucoseconcentration. Subject salivary samples were measured prior toadministration and at 30 mins following administration. The liquidcomposition was ingested in this example as the composition is non-toxicand meant for consumption. Testing was performed using test strips(Multistix, Siemens). Differences in pH, protein content and glucoselevels were observed in all subjects.

An example of results obtained from oral administration of rapid melttablet compositions (ranging from about 320 to about 350 mgs per tablet)comprising filler (in some non-limiting examples, xylitol, sorbitol,Stevia or mannitol was used, ranging from about 310 to about 345 mgs pertablet), chelator (alpha-cyclodextrin, ranging from about 5 to about 30μg per table), base (pyridoxal-5-phosphate, ranging from about 0.5 toabout 15 μg per tablet), enhancer (sodium bicarbonate, ranging fromabout 0.3 to about 12 μg per tablet), and flavoring agent (peppermint)to human subjects is presented in FIG. 4B. Subject salivary samples wereassessed for pH using pH test strip prior to tablet administration andat 30 mins following tablet administration. Differences in pH wereobserved in all subjects. These data teach that formulations comprisingchelator and base compositions can be administered to affect microbialmetabolism, reduce pathology-inducing effects and promote oral health.

Example 5. Compositions to Selectively Affect Pathogens Over Commensals

Compositions comprising chelator and base can selectively affectpathogens over commensal microbes of a mammal's microbiota. The adulthuman gut contains up to 100 trillion microbial organisms and allmucosal and dermal surfaces of mammals comprise microbes, known as themicrobiota of a mammalian host. Specific strains of bacteria have beenimplicated in the disease pathogenesis of cancer, oral diseases,cardiovascular diseases, diseases of the central nervous system,metabolic diseases and systemic diseases. Streptococcus, Bacteriodes,Clostridia, and Helicobacter species are a few of many microbes havingbeen implicated in disease. Conversely, some microbes, includingLactobacillus acidophilus and Bifidobacterium longum, have been shown tobe protective against certain diseases. Thus, the balance between“detrimental” microbes (referred to herein as pathogens) and“beneficial” microbes (referred to herein as commensals) may haveimplications for disease risk, progression and/or severity in mammals.Non-limiting examples of select chelator and base agents that can beused in compositions to selectively affect pathogens without adverselyaffecting commensals are presented in FIG. 5.

Example 6. Health-Promoting Compositions and Formulations

Compositions comprising chelator and base can be administered forpromoting and maintaining health. Compositions may be formulated in asolid dosing form such as tablet, capsule, powder, crystalline, gum,thin strip, lozenge, patch, or other. Solid dosing formulations maycomprise plasticizer, polymer, filling agent, sweetening agent, coloringagent or flavoring agent. Compositions may be formulated in time-releasedosing forms.

Solid dosing formulations may be administered frequently or on an asneeded basis and may be packaged in a single dose, a multi-dose pack, orother. Formulations may be administered orally for sublingual absorptionor may be ingested for absorption in the stomach or intestine.Formulations may also be administered dermally in a patch for absorptionby the skin.

Compositions may also be formulated in a liquid dosing form such asliquid, solution, suspension, emulsion, gel, crème, lotion, serum,elixir, or other. Liquid dosing formulations may comprise plasticizer,polymer, filler, sweetening agent, coloring agent, flavoring agent,fragrance, inhalant, evaporant or other. Compositions may be formulatedin time-release dosing forms and may comprise nanoparticles,microparticles, or crystals.

Liquid dosing formulations may be administered frequently or on an asneeded basis and may be packaged in a single dose, multi-dose pack,concentrate or other. Formulations may be administered: for oralabsorption in the mouth or gut; absorption by the skin; aerosolized ornebulized for absorption by the nasal and sinus cavities or by thelungs; rectally for absorption by the gut; for absorption by the eye; inthe genitourinary tract; or for absorption by the ear.

Example 7. Administration of Compositions Following Consumption ofSugar- and CID-Containing Foods and Beverages

Compositions comprising chelator and base can be administered orally forpromoting and maintaining health following consumption of foods andbeverages that promote microbial growth, undesirable microbialmetabolism, or inflammation. Compositions may be formulated in a directdose or may be formulated in a time-release formula. Compositions may beformulated in liquid or solid dose forms for sublingual absorption orabsorption by the stomach or intestine. Compositions may be administeredon an as needed basis following consumption of food or beverage.

Example 8. Compositions for Topical Administration

Compositions comprising chelator and base can be used for topicaladministration to affect microbial biofilm, microbe-specific metabolismand inflammation of the skin. Human skin is a complex organ thatprotects and regulates the interaction with the external environment.Microbial biofilms permanently inhabit the epidermis with transientmicrobes that occur through contact-mediated transmission. The topicalmicrobial flora is composed primarily of Gram-positive bacteria andfungus. Topical infections often result from injury to the skin, whichprovides access of microbial biofilms to the underlying tissue. Largebacterial populations in wounds have been correlated with delayedhealing, and control of microbial infection is recognized as animportant aspect of wound care (Seth et al. (2012) PLoS One 7:e42897).

Skin-residing microbial biofilms have also been strongly associated withacne and dandruff. Propionibacterium acnes and Staphylococcusepidermidis, in addition to approximately 16 other bacterial speciesconstitute the follicular microbiome (Bek-Thomsen et al. (2008) J ClinMicrobiol. 46:3355-3360). Over 85% of the population has been afflictedby this disease, which accounts for more than 14 million acne-relatedclinical visits per year in the United States. Data also suggest thatacne may contribute to significant psychological distress, depressionand even increased risk of suicide in teenagers suffering from severeacne (Mancini A J. (2008) Johns Hopkins Adv Stud in Med. 8:100-105;Hanisah et al. (2009) J Prim Healthcare. 1:20-25).

An example of a composition to limit the fungal burden residing on thehuman scalp is presented in FIG. 7. As exemplified in FIG. 7, fungus wassampled from human scalps using sterile swabs and cultured in Sabouraudliquid media (pH 5.4). Sensitivity to various compositions were testedusing traditional microbiological culture methods (for example,Sabouraud-brain-heart infusion plates supplemented with chloramphenicolto inhibit bacterial growth). Compositions may be selected for one ormore particular fungal species in a personalized manner. In othernon-limiting examples, subjects were treated for 5 mins with controlsolution (10 mM NaCl) or various compositions (50 μL per donor) on theleft wrist. Treatment zones were outlined on each subject by permanentmarker prior to washing with sterile water. Treatment zones were thensampled by sterile swab and compared to sampling the untreated wrist(right wrist). Swabs were plated on Sabouraud-brain-heart infusionplates supplemented with chloramphenicol to inhibit bacterial growth andallowed to grow for 5 days. Fungal growth between treatment and controlzones were assessed and tabulated. Results from these studies may beused to inform selection of a composition with desired effectiveness.Compositions may be formulated with detergents, surfactants, naturaloils, coloring agents, fragrances, polymers, salts, waxes, emulsifiers,antioxidants, and moisturizers.

Compositions for topical administration may be formulated in washes,wipes, make-up, crèmes, lotions, shampoos, conditioners, gels, mousse,sprays, deodorants or other.

Compositions may be applied to skin surfaces, followed by manual rubbingand massaging actions. Compositions may also be contacted to skinsurfaces by spray, followed by manual rubbing and massaging actions. Yetin other methods, compositions may be applied to the skin, followed byremoval of the compositions by rinsing. Topical compositions may beapplied routinely in a single dose format or may be applied periodicallythroughout the day.

In some non-limiting examples, topical compositions may be used toreduce microbial burdens, to reduce microbial metabolic activity, toreduce metabolically-derived malodor on skin surfaces, and to reducemicrobe-triggered inflammation of the skin.

Example 9. Composition for Affecting Malodor

Malodor is considered an aesthetic problem by many people worldwide.Considerable time and money is spent to limit odor due to inherentsocial and cultural biases. Current methods to affect malodor involvethe application of “odor masking” technologies, such as deodorant andperfume. Other methods involve the application of “odor sequestration”technologies such as talcs and powders. Yet other methods involve theapplication of “antiperspirant” creams, pastes, solutions and others toblock sweat and oil secretion by the skin.

Here, we present compositions affecting microbe-mediated malodor.Mucosal and dermal malodor is known to arise from the metabolic activitymicrobes living on the skin and mucosal surfaces of a mammal. Malodor isproduced by aromatic metabolic waste products generated from microbialmetabolism. Commonly, commensal microbes use the secreted oils of theskin as carbon sources for energy production and use consumed food ascarbon sources in the gut of the mammalian host. Aromatic waste productsmay include butyric acid, acetic acid, propionic acid, sulfites,volatile aromatic compounds and other. Chelator and base compositionsmay be formulated in a powder, crème, gel, solid, lotion, wash, gum,lozenge, tablet, capsule, liquid, gel, deodorant or wipe to inhibitmicrobe-mediated odor production. Compositions may be administeredtopically, orally, or rectally by manual application, through the use ofa device such as a wipe. Compositions may be administered frequently ona daily basis or on an as needed basis.

Example 10. Health-Promoting Compositions for Surfaces

Compositions comprising chelator and base can be administered to foodand surfaces to promote health. Pathogen-harboring microbial biofilmcontamination of food is a significant problem worldwide. Foodborneillness is not decreasing in occurrence despite improvements in foodhandling, food processing and food storage practices. Foodborne illnessaccounts for 48 million illnesses, 128,000 hospitalizations and 3,000deaths each year in the United States (USDA). Every year in the US,millions of tons of chicken, beef and pork products are recalled due topotential food safety issues (CDC).

Compositions comprising chelator and base may be used for food safety.In some embodiments, compositions may be administered to the surface offoods to reduce the microbial CFU burden (examples are demonstrated inFIG. 8). In these embodiments, compositions or commercial vegetable washwas administered to fruits and vegetables for about 5 to 15 seconds andrinsed with tap water. Microbial CFU burden was evaluated using standardmicrobiologic techniques. Various chelator and base combinations weresuperior to commercial products and provide a safe, non-toxic andconsumable method for promoting health.

Some compositions may be used as an additive in processed food andbeverage and can be formulated in powders, liquids, gels, concentratesor other. Preferred methods of food safety compositions relate to thecontacting of food and beverage during the course of preparation,processing and/or packaging said food and beverage.

Said food safety compositions may also be used as device surfacetreatments in food and beverage processing and preparation. Methodsusing food safety compositions relate to the administration of saidcomposition to device surfaces that contact food during the course ofprocessing and preparation. Methods may include the spraying of saidcomposition to cutting blocks, conveyor belts, knives, forks, skewers,processing tools, processing machinery, packaging machinery andpackaging materials.

Hospital-acquired (nosocomial) infections from contaminated devices orpathogen-carrying healthcare professionals results in incidentalinfections at a rate of approximately 4-5.5% in in-patient populationsannually. The healthcare costs associated with biofilm contamination areestimated at $4.5 billion USD (Emori and Gaynes (1993) Clin MicrobiolRev. 6:428-442). Numerous approaches have been employed to reduceunnecessary exposure to pathogen-harboring biofilms, however, thereremains an urgent need for novel and cost-effective approaches toprevent biofilm-associated illness through the administration ofanti-microbial sprays, washes and additives.

Compositions comprising chelator and base may be used on biologic andinanimate surfaces as general cleaners to promote health. Exemplarysurfaces may include doorknobs, keyboards, remote controls, cell phones,handles, elevator buttons, floors, counters, faucets, tools and others.Compositions for administration to surfaces may be formulated aspowders, gels, foams and liquids. Methods of administering compositionsrelate to the contacting of surfaces with the composition for adesirable period of time to disinfect, cleanse, or treat contaminatedsurfaces.

Example 11. Method of Testing Followed by the Selection OF DesirableHealth-Promoting Chelator and Base Composition

Test methods to measure indicators of microbe-mediated inflammation,unwanted microbial metabolism, biofilm, or microbial overgrowth in amammal may relate to the use of one or more test to measure the pH,nitrate level, protein content, leukocyte esterase level, cytokinelevel, dextran content of a sample or a surface. Test may be utilized ona routine basis, daily, weekly, monthly or on an as needed basis. Theuse test may aid in the: selection of health-promoting composition,determination of dose, dose form, formulation, frequency of use, routeof administration and effectiveness.

Test methods may include the administration of a dye to stain surfacesor samples suspected of having evidence of microbe-mediatedinflammation, unwanted microbial metabolism, biofilm, or microbialovergrowth. Dye stains may comprise commercial formulations includingbut not limited to dextran-binding dyes, hematoxylin and eosin,immunohistochemical stains, nitrite- or nitrate-sensitive dyes, proteindyes, sulfur-sensitive dyes and other. Test methods may also include theuse of a test strip, lateral flow test, standard sandwich immunoassay,flow cytometric or other methods.

The examples, which are intended to be purely exemplary of the inventionand should therefore not be considered to limit the invention in anyway, also describe and detail aspects and embodiments of the inventiondiscussed above. The foregoing examples and detailed description areoffered by way of illustration and not by way of limitation. Althoughthe foregoing invention has been described in some detail by way ofillustration and example for purposes of clarity of understanding, itwill be readily apparent to those of ordinary skill in the art in lightof the teachings of this invention that certain changes andmodifications may be made thereto without departing from the spirit orscope of the appended claims.

What is claimed is:
 1. A solid dosage form comprising a cyclodextrin andbase, wherein the cyclodextrin and base are active ingredients thatreduce pathogenic microbial activity, and wherein the base comprisespyridoxal-5-phosphate and is present at about 50 ng to about 1,000 mgper dose, and wherein the cyclodextrin is alpha, beta, or gammacyclodextrin and is present at about 50 ng to about 1,000 mg per dose.2. The solid dosage form of claim 1, wherein the amount of thecyclodextrin is between 0.0005% and 30% of the solid dosage form.
 3. Thesolid dosage form of claim 1, wherein the amount of base is between0.0001% and 15% of the solid dosage form.
 4. The solid dosage form ofclaim 1, further comprising an enhancer.
 5. The solid dosage form ofclaim 4, wherein the enhancer is proline, phenylalanine, boric acid,ascorbic acid, sugar alcohols, or extracts from citrus, berries, teas,peppermint, mint or cinnamon and the amount is between 0.0001% and 10%of the solid dosage form.
 6. The solid dosage form of claim 1, whereinthe solid dosage form is for oral care.
 7. The solid dosage form ofclaim 1, wherein the solid dosage form is formulated in a tablet,powder, paste, deodorant, anti-perspirant, capsule, rapid melt tablet,thin strip, or gum.
 8. The solid dosage form of claim 1, wherein thesolid dosage form is formulated for administration at least once perday.
 9. The solid dosage form of claim 1, wherein the solid dosage formis used to promote oral health.
 10. The solid dosage form of claim 1,wherein the solid dosage form is for use following consumption of aphosphoric acid or ascorbic acid containing beverage.
 11. The soliddosage form of claim 1, wherein the solid dosage form is for usefollowing consumption of food.
 12. A method of promoting oral health ina mammal comprising: identifying an individual at increased risk fordeveloping or exhibiting signs of oral disease; measuring pH, leukocyteesterase, nitrate, microbial marker levels, or plaque-staining dyeretention in the mouth of the individual; comparing said levels tohealthy levels to determine if oral pathology is present; and contactingan effective amount of a solid dosage form comprising a cyclodextrin andbase, to an oral surface if evidence of oral pathology is present,wherein the cyclodextrin and base are active ingredients that reducepathogenic microbial activity, and wherein the base comprisespyridoxal-5-phosphate and is present at about 50 ng to about 1,000 mgper dose, and wherein the cyclodextrin is alpha, beta, or gammacyclodextrin and is present at about 50 ng to about 1,000 mg per dose.13. The solid dosage form of claim 1, wherein the cyclodextrin and baseare formulated for contact with a biological surface.
 14. The soliddosage form of claim 1, further comprising arginine or a polyarginine.15. The solid dosage form of claim 1, wherein the solid dosage form hasa pH range of approximately 4.2 to 7.2.
 16. A method of reducingpathogenic microbial activity, the method comprising: contacting aneffective amount of a solid dosage form comprising a cyclodextrin andbase with a surface, wherein the cyclodextrin and base are activeingredients that reduce pathogenic microbial activity, and wherein thebase comprises pyridoxal-5-phosphate and is present at about 50 ng toabout 1,000 mg per dose, and wherein the cyclodextrin is alpha, beta, orgamma cyclodextrin and is present at about 50 ng to about 1,000 mg perdose.
 17. The method of claim 16, wherein the surface is a biologicalsurface.
 18. The method of claim 17, wherein the biological surface isselected from the group consisting of food, dermal surfaces, toothsurfaces, and mucosal surfaces.
 19. The method of claim 16, wherein thesurface is an inanimate surface.
 20. The method of claim 19, wherein theinanimate surface is selected from the group consisting of floors,counters, utensils, tools, and handles.
 21. The method of claim 16,wherein the base further comprises sodium bicarbonate, arginine, or acombination thereof.
 22. The method of claim 16, wherein the soliddosage form further comprises an enhancer.
 23. The method claim 22,wherein the enhancer is proline, phenylalanine, boric acid, ascorbicacid, sugar alcohols, or extracts from citrus, berries, teas,peppermint, mint or cinnamon and the amount is between 0.0001% and 10%of the solid composition.
 24. The method of claim 22, wherein the basefurther comprises sodium bicarbonate, arginine, or a combinationthereof.
 25. A method of promoting oral health in an individualcomprising administering an effective amount of a solid dosage formcomprising a cyclodextrin and base to the individual, wherein thecyclodextrin and base are active ingredients that reduce pathogenicmicrobial activity, and wherein the base comprises pyridoxal-5-phosphateand is present at about 50 ng to about 1,000 mg per dose, and whereinthe cyclodextrin is alpha, beta, or gamma cyclodextrin and is present atabout 50 ng to about 1,000 mg per dose.
 26. The method of claim 25,wherein the base further comprises sodium bicarbonate, arginine, or acombination thereof.
 27. The method of claim 25, wherein the individualhas one or more of halitosis, tooth plaque, tooth decay, periodontitis,gingivitis and cavity.
 28. The method of claim 25, wherein the soliddosage form further comprises an enhancer.
 29. The method of claim 28,wherein the enhancer is proline, phenylalanine, boric acid, ascorbicacid, sugar alcohols, or extracts from citrus, berries, teas,peppermint, mint or cinnamon and the amount of the enhancer is between0.0001% and 10% of the solid dosage form.
 30. The method of claim 28,wherein the base further comprises sodium bicarbonate, arginine, or acombination thereof.
 31. The method of claim 25, wherein the soliddosage form has a pH range of approximately 4.2 to 7.2.
 32. The methodof claim 25, wherein the solid dosage form has a pH range ofapproximately 6 to
 9. 33. The method of claim 25, wherein the soliddosage form is administered following consumption of a phosphoric acidor ascorbic acid containing beverage.
 34. The method of claim 25,wherein the solid dosage form is administered following consumption offood.
 35. The solid dosage form of claim 1, wherein the cyclodextrin andbase are formulated for contact with an inanimate surface.
 36. The soliddosage form of claim 35, wherein the inanimate surface is selected fromthe group consisting of floors, counters, utensils, tools, and handles.37. The solid dosage form of claim 13, wherein the biological surface isselected from the group consisting of food, dermal surfaces, toothsurfaces, and mucosal surfaces.
 38. The solid dosage form of claim 1,wherein the base further comprises sodium bicarbonate, arginine, or acombination thereof.
 39. The solid dosage form of claim 5, wherein thebase further comprises sodium bicarbonate, arginine, or a combinationthereof.
 40. The solid dosage form of claim 1, wherein the solid dosageform has a pH range of approximately 6 to
 9. 41. A liquid compositioncomprising a cyclodextrin and base, wherein the cyclodextrin and baseare active ingredients that reduce pathogenic microbial activity, andwherein the base comprises pyridoxal-5-phosphate and has a concentrationof about 750 pM to about 250 mM, and wherein the cyclodextrin is alpha,beta, or gamma cyclodextrin and has a concentration of about 0.1 nM toabout 500 mM.
 42. The liquid composition of claim 41, wherein the basefurther comprises sodium bicarbonate, arginine, or a combinationthereof.
 43. The liquid composition of claim 41, wherein theconcentration of the cyclodextrin is between 0.0005% and 30% of theliquid composition.
 44. The liquid composition of claim 41, wherein theconcentration of base is between 0.0001% and 15% of the liquidcomposition.
 45. The liquid composition of claim 41, further comprisingan enhancer.
 46. The liquid composition of claim 45, wherein theenhancer is proline, phenylalanine, boric acid, ascorbic acid, sugaralcohols, or extracts from citrus, berries, teas, peppermint, mint orcinnamon and the concentration is between 0.0001% and 10% of the liquidcomposition.
 47. The liquid composition of claim 45, wherein the basefurther comprises sodium bicarbonate, arginine, or a combinationthereof.
 48. The liquid composition of claim 41, wherein the liquidcomposition is for oral care.
 49. The liquid composition of claim 41,wherein the liquid composition is formulated as a spray, deodorant,anti-perspirant, cream, gel, concentrate, wipe, or wash.
 50. The liquidcomposition of claim 49, wherein the wash is a mouthwash.
 51. The liquidcomposition of claim 41, wherein the liquid composition has a pH rangeof approximately 4.2 to 7.2.
 52. The liquid composition of claim 41,wherein the liquid composition has a pH range of approximately 6 to 9.53. The liquid composition of claim 41, wherein the cyclodextrin andbase are formulated for contact with a biological surface.
 54. Theliquid composition of claim 53, wherein the biological surface isselected from the group consisting of food, dermal surfaces, toothsurfaces, and mucosal surfaces.
 55. The liquid composition of claim 41,wherein the cyclodextrin and base are formulated for contact with aninanimate surface.
 56. The liquid composition of claim 55, wherein theinanimate surface is selected from the group consisting of floors,counters, utensils, tools, and handles.
 57. A method of reducingpathogenic microbial activity, the method comprising contacting aneffective amount of a liquid composition comprising a cyclodextrin andbase with a surface, wherein the cyclodextrin and base are activeingredients that reduce pathogenic microbial activity, and wherein thebase comprises pyridoxal-5-phosphate and has a concentration of about750 pM to about 250 mM, and wherein the cyclodextrin is alpha, beta, orgamma cyclodextrin and has a concentration of about 0.1 nM to about 500mM.
 58. The method of claim 57, wherein the surface is a biologicalsurface.
 59. The method of claim 58, wherein the biological surface isselected from the group consisting of food, dermal surfaces, toothsurfaces, and mucosal surfaces.
 60. The method of claim 57, wherein thesurface is an inanimate surface.
 61. The method of claim 60, wherein theinanimate surface is selected from the group consisting of floors,counters, utensils, tools, and handles.
 62. The method of claim 57,wherein the base further comprises sodium bicarbonate, arginine, or acombination thereof.
 63. The method of claim 57, wherein the liquidcomposition further comprises an enhancer.
 64. The method of claim 63,wherein the enhancer is proline, phenylalanine, boric acid, ascorbicacid, sugar alcohols, or extracts from citrus, berries, teas,peppermint, mint or cinnamon and the concentration is between 0.0001%and 10% of the liquid composition.
 65. The method of claim 63, whereinthe base further comprises sodium bicarbonate, arginine, or acombination thereof.
 66. A method of promoting oral health in anindividual comprising administering an effective amount of a liquidcomposition comprising a cyclodextrin and base to the individual,wherein the cyclodextrin and base are active ingredients that reducepathogenic microbial activity, and wherein the base comprisespyridoxal-5-phosphate and has a concentration of about 750 pM to about250 mM, and wherein the cyclodextrin is alpha, beta, or gammacyclodextrin and has a concentration of about 0.1 nM to about 500 mM.67. The method of claim 66, wherein the base further comprises sodiumbicarbonate, arginine, or a combination thereof.
 68. The method of claim66, wherein the individual has one or more of halitosis, tooth plaque,tooth decay, periodontitis, gingivitis and cavity.
 69. The method ofclaim 66, wherein the liquid composition further comprises an enhancer.70. The method of claim 69, wherein the enhancer is proline,phenylalanine, boric acid, ascorbic acid, sugar alcohols, or extractsfrom citrus, berries, teas, peppermint, mint or cinnamon and theconcentration is between 0.0001% and 10% of the liquid composition. 71.The method of claim 69, wherein the base further comprises sodiumbicarbonate, arginine, or a combination thereof.
 72. The method of claim66, wherein the liquid composition has a pH range of approximately 4.2to 7.2.
 73. The method of claim 66, wherein the liquid composition has apH range of approximately 6 to
 9. 74. The method of claim 66, whereinthe liquid composition is administered following consumption of aphosphoric acid or ascorbic acid containing beverage.
 75. The method ofclaim 66, wherein the liquid composition is administered followingconsumption of food.
 76. A method of promoting oral health in a mammalcomprising: identifying an individual at increased risk for developingor exhibiting signs of oral disease; measuring pH, leukocyte esterase,nitrate, microbial marker levels, or plaque-staining dye retention inthe mouth of the individual; comparing said levels to healthy levels todetermine if oral pathology is present; and contacting an effectiveamount of a liquid composition comprising a cyclodextrin and base to anoral surface if evidence of oral pathology is present, and wherein thecyclodextrin and base are active ingredients that reduce pathogenicmicrobial activity, wherein the base comprises pyridoxal-5-phosphate andhas a concentration of about 750 pM to about 250 mM, and wherein thecyclodextrin is alpha, beta, or gamma cyclodextrin and has aconcentration of about 0.1 nM to about 500 mM.